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"Terhambatnya perkembangan otak dan saraf merupakan problem kesehatan, diperkirakan mencapai 17% dari seluruh populasi. Dampaknya dapat menurunkan fungsi kognitif/daya ingat. Dasar patologi penurunan fungsi kognitif antara lain disebabkan oleh berkurangnya sinaps, neuron, neurotransmitter dan jejaring saraf. Hal ini berkaitan dengan sinyal-sinyal penting seperti faktor neurotrofik, neurotransmitter dan hormon. Pegagan (Centella asiatica) telah lama digunakan secara empiris untuk memperbaiki daya ingat. Hasil penelitian secara in vivo pemberian pegagan dapat meningkatkan level GABA(Gamma aminobutyric acid) di otak. Pengaruh pegagan pada BDNF(Brain derived neurotrophic factor) belum pemah diteliti, namun menurut Obrietan dkk stimulasi GABA dapat meningkatkan ekspresi BDNF melalui jalur MAPK-CREB (mirogen ocrivated prorein kinase-cyclic AMP response element binding protein). Brain derived neurotrophic factor (BDNF) merupakani salah satu substansi dalam pengaturan neurogenesis. Penelitian ini merupakan studi eksperimental untuk meneliti kadar BDNF dan jumlah sel saraf pada kultur jaringan hipokampus tikus muda. Pengukuran kadar BDNF dengan spektrofotometer pada panjang gelombang 450 nm dengan kit BDNF dari Chemicon. Data dianalisis dengan ANOVA (anabrsis of varions), dan sebelumnya diuji normalitas data dengan Lavene, serta post Hoc Test. Dari penelitian ini diperoleh hasil (1) jumlah sel saraf lebih besar pada kultur sel jaringan hipokampus tikus muda yang diberi ekstrak pegagan 0,50 ug/ml dibandingkan 0,25 ug/ml dan kontrol sebagai pembanding (p<0,05). (2) Untuk kadar BDNF terlihat hasil kadar BDNF pada kelompok kontrol lebih tinggi dibandingkan perlakuan ekstrak pegagan 0,25 ug/ml dan 0,50 ug/ml (p>0,05)."

"ABSTRAKLatar belakang. Proses inflamasi kronik dan persisten mempengaruhi tingginya rekurensi dan survival endometriosis pasca pembedahan. Hal ini menjadi permasalahan endometriosis, sehingga perlu pengembangan terapi target salah satunya yaitu asam galat. Asam galat terbukti efektif sebagai antikanker, anti tumor, anti inflamasi dan antibakterial pada beberapa cell line, namun efektifitasnya pada sel endometriosis harus dibuktikan. Tujuan. membuktikan efek asam galat dan senyawa turunannya terhadap regulasi inflamasi pada kultur primer endometriosis ditinjau dari ekspresi mRNA NF-kB, serta sekresi TNF-? dan IL-6. Metode. Sel endometriosis berasal dari jaringan endometriosis pasien yang menjalani laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil dan oktil galat dengan dosis 25,6 g/mL, 51,2 g/mL dan 102,4 g/mL selama 48 jam, kemudian diinduksi dengan LPS 500 ng/mL selama 24 jam. Regulasi inflamasi dinilai dari ekspresi mRNA NF-kB dengan qRT-PCR, kadar sekresi TNF-? dan IL-6 dengan ELISA, serta inhibisi viabilitas sel dengan MTS Assay. Hasil. Setelah data dirasiokan dengan kontrol, ketiga zat signifikan menghambat viabilitas sel endometriosis p value 0,000 dengan inhibisi tertinggi pada dosis 102,4 g/mL. Terjadi penurunan ekspresi relatif NF-kB yang dirasiokan dengan kontrol dan IL-6 meskipun secara statistik tidak bermakna. Konsentrasi TNF? tidak berbeda secara bermakna p value 0,340 . Kesimpulan. Asam galat dan senyawa turunannya berpengaruh terhadap inhibisi viabilitas sel, penurunan ekspresi relatif NF-kB dan IL-6, namun tidak bermakna terhadap penekanan sitokin TNF-?. Perlu dilakukan studi lanjut untuk menilai efektifitas asam galat sebagai kandidat obat antiinflamasi pada endometriosis ditinjau aspek lain.

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ABSTRACTBackground. Endometriosis is a benign gynecological disorder characterized by the growth of the lining of the endometrium like tissue outside the uterus. The cause of the growth of endometriosis is not known well, chronic and persistent inflammatory process is suspected to be one of the pathogenesis that contributes to the high recurrence and survival endometriosis. One of the potential therapeutic agents is a gallic acid which proved effective in earlier studies as an anti cancer, anti tumor, anti inflammatory and antibacterial in several cell line. The Effectiveness of gallic acid to the endometriosis cell is a preliminary study and have not found evidence of publication yet. Object. Proving the effect of gallic acid and its derivatives on the inflammatory regulation of endometriosis primary culture study on mRNA expression of NF kB, TNF , and IL 6 secretion. Method. Endometriosis cells from Indonesian endometriosis patients tissues who had undergone laparoscopy surgery were isolated by the enzymatic reaction and primary cultured. Cultured cells treated by gallic acid and alkyl ester synthetic derivatives of the gallic acids heptyl gallate and octyl gallate each with the dosage of 25,6 g mL, 51,2 g mL, and 102.4 g mL for 48 hours and then induced by LPS 500 ng mL for 24 hours. Parameter research was assessed by qRT PCR for mRNA expression of NF kB, ELISA for the quantification of TNF and interleukin 6, and MTS assay was used to observe endometriosis cell viability. Results. After the data was rationalized with the control, three substances showed significant inhibition of endometriosis cell viability. The highest inhibition for all treatment was at doses 102,4 g mL. Overall there was an inhibition of relative expression of mRNA NF kB were rationalized to controls and suppression of IL 6 in octyl gallate groups. The concentration of TNF among the groups did not differ significantly p value 0.340 . Conclusion. Gallic acid and its derivatives have significantly inhibition effect toward cell viability, mRNA expression of NF kB, and IL 6 but have not significantly effect toward cytokine TNF . Further studies need to be conducted to assess the effectiveness of gallic acid as an anti inflammatory drug candidate toward to any pathway."

"ABSTRAKLatar belakang. Potensi terjadinya kekambuhan paska pengobatan endometriosisdengan terapi hormonal dan pembedahan konservatif masih terjadi sekitar 11-32dalam waktu 1-5 tahun. Salah satu faktor pemicunya adalah proses inflamasi kronikyang merangsang peningkatan sitokin proinflamasi dalam rongga peritoneum, sehinggaperlu pengembangan terapi baru. Heptil galat dan oktil galat merupakan senyawaturunan asam galat yang berpotensi menekan proliferasi beberapa jenis sel kanker.Penelitian kami sebelumnya membuktikan oktil galat dapat menekan ekspresi mRNANFkB yang merupakan faktor transkripsi aktivasi jalur proinflamasi, serta dapatmenekan proliferasi sel endometriosis in vitro. Saat ini kami ingin menganalisisaktivitas heptil galat dan oktil galat terhadap protein target NFkB melalui teknik insilicodocking dan efeknya terhadap regulasi sitokin proinflamasi IL-1, COX-2, TGF-1 dan IL-10 pada kultur primer sel endometriosis.Metode. In silico docking heptil galat dan oktil galat terhadap protein target NFkBmelalui teknik bioinformatika. Sel endometriosis dari jaringan primer pasien diisolasisecara enzimatis dan dikultur, kemudian diberi perlakuan heptil dan oktil galat dengan 2macam dosis (51,2 μg/mL dan 102,4 μg/mL) selama 48 jam, dilanjutkan induksi LPS 10ng/mL selama 24 jam. Kelompok kontrol positif hanya diinduksi LPS tanpa perlakuan,dan kontrol negatif tanpa perlakuan dan LPS. Regulasi inflamasi dinilai dari tingkatkadar sitokin IL-1, COX-2, TGF-1 dan IL-10 dengan teknik ELISA.Hasil. Analisis in-silico docking protein NFkB menunjukan nilai ikatan energi oktilgalat lebih tinggi (-7,98 kkal/mol) dibandingkan heptil galat (-7,68 kkal/mol) dan asamgalat (-7,66 kkal/mol). Terjadi penurunan kadar sitokin COX-2 secara signifikan(p<0,03) pada kelompok perlakuan dibandingkan dengan kontrol positif, begitu jugadengan sitokin IL-1 dan IL-10 cenderung menurun (p>0,05). Sedangkan kadar sitokinTGF-1 mengalami kenaikan pada kelompok perlakuan dibandingkan kontrol positifmeskipun kurang bermakna secara statistik (p>0,05).Kesimpulan. Melalui jalur NF-kB sebagai regulator inflamasi, baik oktil galat danheptil galat terbukti dapat menekan produksi sitokin proinflamasi COX2 dan IL-1serta meningkatkan sitokin TGF-1 dan menurunkan sitokin IL-10 sehingga berpotensisebagai bahan terapi tambahan pada endometriosis.

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ABSTRACTBackground: The potential for relapse post endometriosis treatment with hormonaltherapy and conservative surgery still occurs around 11-32 within 1-5 years. One ofthe trigger factors is a chronic inflammatory process that stimulates an increaseproinflammatory cytokines in the peritoneal cavity, so needed the development of newtherapies. Heptyl galate and octyl galate are gallic acid derivatives which have thepotential to suppress the proliferation of several types cancer cells. Our previousresearch proved that octyl galate can suppress the expression of NFkB mRNA which isa proinflammatory activation transcription factor, and can suppress endometriosis cellproliferation in vitro. We currently want to analyze the activity of heptyl galates andoctyl galates against the NFκB target protein through in-silico docking techniques andtheir effects on the regulation of proinflammatory cytokines IL-1, COX-2, TGF-1 andIL-10 in primary cultures of endometriosis cells.Method: In silico docking heptyl and octyl galates against the NFkB target proteinsthrough bioinformatics techniques. Endometriosis cells from primary tissue wereenzymatically isolated and cultured, then given heptyl and octyl gallate treatment with 2doses (51.2 μg/mL and 102,4 μg/mL) for 48 hours, continued induction of 10 ng / mLLPS for 24 hours. The positive control group only induced LPS without treatment, andnegative treatment without treatment and LPS. Inflammatory regulation was assessedfrom levels of cytokines IL-1, COX-2, TGF-1 dan IL-10 with ELISA techniques.Results: In-silico docking analysis of the NFkB gene showed higher energy bondingvalues in octyl galate (-7,98 kcal / mol) than heptyl galate (-7,68 kcal / mol) and gallicacid (-7,66 kcal / mol). Significantly decreased levels of COX-2 cytokine (p <0,03) inthe treatment group compared with positive controls, so also the cytokines of IL-1 andIL-10 tended to decrease (p> 0,05). Whereas the levels of cytokine TGF-1 experiencedan increase in the treatment group compared to the positive control although it was lessstatistically significant (p> 0,05).Conclusion: Through the NFkB pathway as an inflammatory regulator, both octylgalates and heptyl galates have been shown to suppress the production ofproinflammatory cytokines COX2 and IL-1, as well as increase TGF-1 cytokines andreduce IL-10 cytokines so that they have the potential to be additional therapeuticagents in endometriosis."

"ABSTRAKEndometriosis merupakan penyakit ginekologi ditandai dengan implantasi jaringan endometrium di luar rongga uterus, berhubungan erat dengan proses inflamasi kronis. Stres oksidatif menjadi aktivator terjadinya proses inflamasi kronis di endometriosis. Oktil galat terbukti lebih efektif menekan proses inflamasi dibandingkan asam galat dan heptil galat pada sel kultur primer endometriosis. Penelitian ini bertujuan menganalisis pengaruh oktil galat pada proses inflamasi dan stres oksidatif pada tikus Wistar model endometriosis. Tiga puluh ekor tikus Wistar dibagi menjadi tiga kelompok, yaitu kelompok uji, kontrol endometriosis dan kelompok normal. Kelompok uji dilakukan autotransplantasi lalu diberikan suspensi oktil galat dan CMC selama satu bulan. Kelompok endometriosis dilakukan autotransplantasi lalu diberikan larutan CMC selama satu bulan, sedangkan kelompok normal hanya dilakukan laparotomi. Seluruh tikus kemudian dieuthanasia, dari kelompok uji dan kontrol endometriosis diambil jaringan endometriosisnya sedangkan dari kelompok sehat diambil jaringan endometriumnya untuk dianalisis. Analisis MDA (Malondialdhyde) dan SOD (Superoxide Dismutase) dilakukan secara spektofotometri, kadar NF-ĸB dengan ELISA dan IL-1β (Interleukin-1 Beta) dengan LUMINEX. Pemberian oktil galat pada kelompok uji tidak menurunkan kadar MDA namun berpotensi menekan kondisi stres oksidatif dengan meningkatkan kadar SOD. Oktil galat terbukti menekan aktivasi NF-ĸB secara signifikan, namun tidak menekan kadar IL-1β. Oktil galat berperan sebagai antiinflamasi pada tikus Wistar model endometriosis dengan cara induksi peningkatan SOD dan hambatan langsung pada translokasi nuklear NF-ĸB.

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ABSTRACTEndometriosis is a gynecological disease characterized by the implantation of endometrial tissue outside the uterine cavity, related to the chronic inflammatory process. Oxidative stress activates the occurrence of chronic inflammatory in endometriosis. Octyl gallate is more effective in suppressing the inflammatory process than gallic acid and heptil gallate in primary endometriosis culture cells. This study aimed to analyze the effect of octyl gallate on the inflammatory process and oxidative stress in endometriosis Wistar rat models. 30 Wistar rats were divided into three groups, the test group, endometriosis control and normal groups. The test group was autotransplantated and then given a suspension of octyl galate and CMC for one month. The endometriosis group was autotransplanted and then given a CMC solution for one month, while the normal group only underwent laparotomy. All rats were then euthanized, from the test and endometriosis group the endometriosis tissue was taken while from the normal group endometrial tissue was taken for analysis. MDA and SOD were measured using spectrophotometry, NF-ĸB with ELISA and IL-1β with LUMINEX. Induction of octyl gallate in the test group did not reduce MDA levels but could potentially suppress oxidative stress conditions by increasing SOD levels. Octyl gallate significantly inhibit the NF-ĸB activation, but not suppressing IL-1β levels significantly. Octyl gallate act as anti-inflammatory agent in endometriosis Wistar rat model through the enhancement of SOD and direct inhibition to nuclear translocation of NF-ĸB."

"Latar Belakang. Endometriosis ditandai dengan pertumbuhan jaringan endometrium di luar uterus, salah satunya disebabkan oleh disregulasi apoptosis sel yang memicu ketahanan sel ektopik. Asam galat dan turunannya pada beberapa penelitian mampu menghambat karsinogenesis pada beberapa cell line kanker. Penelitian kami terdahulu membuktikan asam galat dan turunannya dapat menekan proliferasi sel dan meningkatkan apoptosis sel endometriosis in vitro, namun efeknya terhadap mekanisme jalur apoptosis instrinsik belum di buktikan. Metode. Sel endometriosis berasal dari jaringan endometrium pasien laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil galat, oktil galat dengan dosis 51,2 μg/ml, 102,4 μg/ml dan 153,6 μg/ml selama 48 jam, dilanjutkan induksi 10 ng/ml LPS selama 24 jam . Kelompok kontrol hanya di induksi LPS tanpa perlakuan. Ekspresi relatif mRNA Bax, Bcl-2, dan Caspase-3 dinilai dengan qRT-PCR. Hasil. Peningkatan tertinggi ekspresi mRNA Bax dan penekanan tertinggi ekspresi mRNA Bcl-2 pada oktil galat dosis 153,6 μg/ml. Peningkatan ekspresi mRNA Bax, dan penurunan ekspresi mRNA Bcl-2 akan di ikuti dengan peningkatan ekspresi mRNA Caspase-3. Secara statistik tidak terdapat perbedaan bermakna antara kelompok perlakuan dengan ekspresi mRNA Bax, Bcl-2, dan Caspase-3 (p > 0,05). Kesimpulan. Oktil galat, asam galat, dan heptil galat memiliki efek potensial pada mekanisme apoptosis intrinsik.

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Background. Endomeriosis characterized by the presence of extrauterine endometrial tissue, one of which caused by disregulation of apoptosis that contribute of endometrial ectopic survival. Our previous research has proven that gallic acid and its derivatives can suppress proliferation and induce apoptosis endometriosis cell in vitro. However, the effect of gallic acid and its derivatives on apoptosis intrinsic pathway mechanism is not proven yet. Method. Endometriosis cell from endometriosis patiens who had undergone laparascopy surgery were isolated by enzimatic reaction and primary cultured. Cultured cells treated by gallic acid, heptyl gallate and octyl gallate each with dosage 51.2 μg/ml, 102.4 μg/ml, 153.6μg/ml for 48 hours, than induced by LPS 10 ng/ml for 24 hours. Parameter research was assessed by qRT-PCR for mRNA expression of Bax, Bcl-2, Caspase-3. Result. Octyl gallate showed more effect to induce apoptosis intrinsic . Endometriosis cell were treated with octyl gallate shown increases of Bax and Caspase3 mRNA expression than decrease of Bcl-2 mRNA expression. Statistically, mean differences are not significant between treatment groups and mRNA expression (p > 0.05). Conclusion. This study exhibited that octyl gallate has a more potential effect on apoptosis intrinsic in endometriosis cell cultures followed by gallic acid and heptyl gallate and their potency as treatment for endometriosis."

"Mesenchymal Stem cells (MSCs) endometrium dengan marka spesifik SUSD2 berpotensi menjadi salah satu sumber sel yang menginisiasi lesi ektopik pada endometriosis. MSCs pada ektopik (EK) dan eutopik endometrium (EU) pasien endometriosis diketahui memiliki karakteristik yang lebih invasif,, di antaranya dengan ekspresi indoleamine 2,3 dioxygenase (IDO1) yang lebih tinggi dibandingkan endometrium normal (EN). IDO1 menekan ekspresi p53, protein terkait apoptosis, menyebabkan viabilitas sel endometriosis meningkat. Oktil galat merupakan senyawa pleiotropik yang terbukti mampu menginduksi apoptosis pada sel endometriosis melalui pengamatan mikroskop konfokal. Penelitian ini bertujuan untuk mengetahui pengaruh OG terhadap ekspresi SUSD2 dan jalur antiapoptosis sel melalui IDO1. Bahan biologis tersimpan sel endometriosis positif marka MSCs CD73, CD90, dan CD105 digunakan dalam penelitian. Ekspresi protein SUSD2 diinvestigasi dengan flowcytometry. Ekspresi mRNA IDO1 dan caspase3 diinvestigasi dengan real time RT-PCR. Hasil menunjukkan gambaran penurunan ekspresi SUSD2 pasca-pemberian OG. Terdapat gambaran peningkatan mRNA IDO1 tanpa diikuti penurunan mRNA caspase3 pada EK dan EN, namun tidak pada EU. Kesimpulan penelitian ini adalah oktil galat diduga menginduksi apoptosis, tetapi tidak melalui jalur antiapoptosis IDO1.

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Endometrial mesenchymal stem cells (MSCs) with specific marker SUSD2 is potential to be the sources of the cells that initiate ectopic lesions in endometriosis. MSCs in ectopic (EC) and eutopic endometrium (EU) of endometriosis patients were known to have more invasive characteristics, including higher expression of indoleamine 2,3 dioxygenase (IDO1) than normal endometrium (EN). IDO1 suppresses p53 expression, a protein associated with apoptosis, causing increased endometriosis cell viability. Octyl gallate is pleiotropic compound that has been shown to induce apoptosis in endometriosis cells as seen by confocal microscopy observation. This study aims to determine the effect of OG on the expression of SUSD2 and cell antiapoptotic pathway through IDO1. Stored biological material of endometriosis cell with positive MSCs markers CD73, CD90, and CD105 was used in the study. SUSD2 protein expression was investigated by flowcytometry. IDO1 and caspase3 mRNA expression were investigated with real time RT-PCR. Results showed a trend of decreased SUSD2 expression post-OG administration. There was a trend of increased IDO1 mRNA, but was not followed by a trend of decreased caspase3 mRNA in EK and EN. The conclusion of this study is octyl gallate was suspected to induce apoptosis through a mechanism other than IDO1 antiapoptotic pathway."